ABSTRACT
Objective To investigate the regulation effect of hepatocyte nuclear factor (HNF) 1α on the gene expression profile and the signal pathways in HuH7 cells.Methods The expression of HNF1α was increased or decreased in HuH7 cells by Lenti-virus carrying HNF1α or shHNF1α.The expression profile of the cells after treated was examined by microarray technology.The difference expressed gene regulated by HNF1α were screened and the pathway was analyzed with DAVID software and related analysis system.The regulation effect of HNF1α on transforming growth factor (TGF)β signal pathway was detected by reporter gene test and the regulation role of HNF1α on related genes of TGFβ signal pathway was determined by real-time polymerase chain reaction (PCR) and Western blotting assay.Results The expression of HNF1α in HuH7 cells was significantly up-regulated by Lenti-virus carrying HNF1α gene (Lenti-HNF1α) and which was down regulated by Lenti-virus with shHNF1α gene (LentishHNF1 α).Expression profile analysis revealed that 339 genes were positively up regulated two times by HNF1α and 325 genes were negatively down regulated two times.Signal pathway analysis revealed that HNF1α regulated drug metabolism,biosynthesis of unsaturated fatty acids and glycolysis/gluconeogenesis metabolism signal pathways.Moreover,it also involved in the regulation of TGFβ、nuclear factor (NF)-κB and p53 tumor-related signal pathways.Furthermore,Luciferase reportor gene experiment indicated that up-regulated HNF1α could inhibit the activation of TGFβ signal pathway.And the results of real-time PCR and Western blotting verified that up-regulated HNF1α could inhibit TGFβ signal pathway related gene c-myc and TGFβ1 and then inhibited the activation of TGFβ signal pathway.Conclusion HNF1α broadly affects the gene expression profile and the tumor genesis and development related signal pathways in HuH7 cells,furthermore,HNF1α can inhibit the activation of TGFβ signal pathway.
ABSTRACT
Objective To study the best processing technology of Xanthii Fructus by determining the contents of chlorogenic acid and 3,5-dicaffeoylquinic acid in which processed by different temperature and time. Methods Sixteen batchs samples of Xanthii Fructus were propressed by stir-frying with sand, and the propressed temperature and time were set at 150-220 ℃ and 0.5-7 minutes. Two phenolic acid components in Xanthii Fructus were simultaneously determined. The column was UPLC Acquity BEH C18 (2.1 mm×100 mm, 1.7 μm). The mobile phase was acetonitrile-0.1% phosphoric acid, gradient elution. The flow rate was 0.25 mL/min, and the detection wavelength was 327 nm. Results The sample with highest contents of chlorogenic acid and 3,5-dicaffeoylquinic acid was the batch processed by stir-frying with sand at 160 ℃ for 7 minute, which was 2.498, 2.004 mg/g, respectively. Conclusion According to the appearance of processed sample and the content of chlorogenic acid and 3,5-dicaffeoylquinic acid, the optimal processing technology of Xanthii Fructus was stir-frying with sand at 160 ℃ for 7 min.